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Viruses are obligate intracellular pathogens that are dependent on cellular machineries for their replication. Recent technological breakthroughs have facilitated reliable identification of host factors required for viral infections and better characterization of the virus–host interplay. While these studies have revealed cellular machineries that are uniquely required by individual viruses, accumulating data also indicate the presence of broadly required mechanisms. Among these overlapping cellular functions are components of intracellular membrane trafficking pathways. Here, we review recent discoveries focused on how viruses exploit intracellular membrane trafficking pathways to promote various stages of their life cycle, with an emphasis on cellular factors that are usurped by a broad range of viruses. We describe broadly required components of the endocytic and secretory pathways, the Endosomal Sorting Complexes Required for Transport pathway, and the autophagy pathway. Identification of such overlapping host functions offers new opportunities to develop broad-spectrum host-targeted antiviral strategies.

Trichomonas vaginalis is the etiologic agent of trichomoniasis, one of the most common non-viral sexually transmitted infections globally. Our previous work reported the role of phosphatidylinositol 4,5-bisphosphates (PIP2) signaling in the actin-dependent pathogenicity of T. vaginalis. This study further demonstrated that iron transiently regulated T. vaginalis phosphatidylinositol-4-phosphate 5-kinase (TvPI4P5K) proteostasis and its complex formation with an active ADP ribosylation factor TvArf220, facilitating co-trafficking to the plasma membrane, crucial for PIP2 production. In dominant-active HA-TvArf220 Q71L mutant, TvPI4P5K plasma membrane trafficking, PIP2 production, and intracellular calcium levels were increased, while these processes were inhibited in dominant-negative T31N mutant or those by Brefeldin A (BFA) treatment. Additionally, PIP2 replenishment reversed these inhibitions in the T31N mutant, suggesting the critical role of TvArf220 activation in PIP2-calcium signaling. Also, T31N mutant and BFA treatment impaired actin dynamics and cytoskeleton-dependent processes in T. vaginalis, further linking the role of TvArf220 to PIP2-calcium-dependent actin dynamics. Beyond cytoadherence, during host-parasite interactions, TvArf220 influenced both contact-dependent and -independent cytotoxicity, as well as phagocytotic capacity, contributing to the cytopathogenesis of human vaginal epithelial cells. Our findings underscore the key upstream regulation mechanisms of the PIP2 signaling, orchestrating the interplay between TvArf220-PIP2-calcium signaling and downstream actin cytoskeleton-driven pathogenicity in T. vaginalis.IMPORTANCETrichomonas vaginalis actin cytoskeleton-centric pathogenicity is regulated by the phosphatidylinositol 4,5-bisphosphates (PIP2)-triggered calcium signaling cascade in response to environmental iron, though the detailed mechanism by which iron modulates PIP2 signaling remains unclear. Our findings reveal that iron rapidly induces T. vaginalis phosphatidylinositol-4-phosphate 5-kinase (TvPI4P5K) translation followed by its degradation, while simultaneously activating TvArf220 binding, which facilitates TvPI4P5K localization to the plasma membrane for PIP2 production. In contrast to the TvArf220 Q71L mutant, the reduced PIP2 production, intracellular calcium, actin assembly, morphogenesis, and cytoadherence in the dominant-negative T31N mutant were recovered by PIP2 supplementation, indicating the essential role of TvArf220 in PIP2-dependent calcium signaling. Additionally, the contact-dependent or -independent cytotoxicity, along with the phagocytosis, was impaired in the TvPI4P5K- or TvArf220-deficient parasites, as well as in those treated with BAPTA or Latrunculin B. These findings highlight that TvArf220-mediated PIP2-calcium signaling cascade regulates actin cytoskeleton and cytopathogenicity of T. vaginalis. This study uncovers a novel pathogenic mechanism and suggests potential therapeutic targets for parasite control.

Colorectal cancer (CRC) is a heterogeneous disease with varying clinical outcomes. The identification of distinct subgroups of CRC patients based on molecular profiling can aid in better understanding the disease and improving patient outcomes. This study aimed to investigate the potential of membrane trafficking-related genes (MTRGs) in sub-grouping colorectal cancer patients based on their overall survival and immune microenvironments. Consensus clustering analysis identified two distinct clusters with different expression profiles of membrane trafficking-related genes. The patients in cluster 1 had a significantly better overall survival than those in cluster 2. Furthermore, the immune microenvironments in the two clusters were also found to be significantly different, with cluster 1 having a higher immune score and more immune cells present. Functional analysis of differentially expressed genes between the two clusters revealed that MTRGs were involved in immune response and metabolic processes, and a risk signature model based on MTRGs was established to predict the prognosis of CRC patients. These findings suggest that MTRGs play a crucial role in the immune microenvironment and overall survival of CRC patients and may provide a potential target for personalized therapy.

The role of the primary cilium in key signaling pathways depends on dynamic regulation of ciliary membrane protein composition, yet we know little about the motors or membrane events that regulate ciliary membrane protein trafficking in existing organelles. Recently, we showed that cilium-generated signaling in Chlamydomonas induced rapid, anterograde IFT-independent, cytoplasmic microtubule-dependent redistribution of the membrane polypeptide, SAG1-C65, from the plasma membrane to the periciliary region and the ciliary membrane. Here, we report that the retrograde IFT motor, cytoplasmic dynein 1b, is required in the cytoplasm for this rapid redistribution. Furthermore, signaling-induced trafficking of SAG1-C65 into cilia is unidirectional and the entire complement of cellular SAG1-C65 is shed during signaling and can be recovered in the form of ciliary ectosomes that retain signal-inducing activity. Thus, during signaling, cells regulate ciliary membrane protein composition through cytoplasmic action of the retrograde IFT motor and shedding of ciliary ectosomes. Nearly every cell in the human body has slender, hair-like structures known as cilia that project outwards from its surface. These structures can sense and respond to light, chemicals and touch, and they are required for normal development. Failure of cilia to form or function in the correct manner can lead to severe diseases—such as kidney disorders, deafness and loss of vision. A major puzzle for researchers who study cilia has been to understand how cells change the composition of these structures as part of their response to a sensory input. Cilia are ancient structures that were present in early single-celled organisms and researchers interested in cilia have often used a single-celled green alga called Chlamydomonas reinhardtii as a model system for their studies. When these algae reproduce sexually, the two types of sex cells sense the presence of each other when their cilia touch and then stick together. This ciliary touching activates signals that are sent into the cells to get them ready to fuse together, much like sperm and egg cells do in animals. Both ciliary touching and signaling depend on a protein called SAG1, a part of which (known as SAG1-C65) is normally found mostly over the surface membrane of C. reinhardtii. Only very small amounts of SAG1-C65 are normally found on cilia; but, when the sex cells' cilia touch, this protein rapidly moves to the end of the cell nearest the cilia via a previously unknown mechanism. SAG1-C65 then becomes much more enriched in the cilia. Cao, Ning, Hernandez-Lara et al. investigated this process and found that SAG1-C65 movement requires a molecular motor called ‘cytoplasmic dynein’. This motor protein typically walks along the inside of cilia to transport other molecules away from the tip and towards the cell membrane. However, Cao, Ning, Hernandez-Lara et al. found that this dynein also carries SAG1-C65 from the membrane of the cells towards the base of the cilia in preparation for it to enter into these structures. As part of an effort to understand the fate of the protein after it entered cilia, Cao, Ning, Hernandez-Lara et al. discovered that the SAG1-C65 disappeared from the structures without returning to the cell membrane. Instead, SAG1-C65 was packaged within tiny bubble-like structures near the tips of cilia and these packages were then shed from cilia into the external environment. This discovery challenges a widely held view that proteins are only removed from cilia by returning to the cell. Future work will be required to understand more of the molecular details of these processes, which are likely to be present in most cells with cilia.

[This corrects the article DOI: 10.1371/journal.pone.0130778.].

Pollen tube elongation is characterized by a highly-polarized tip growth process dependent on an efficient vesicular transport system and largely mobilized by actin cytoskeleton. Pollen tubes are an ideal model system to study exocytosis, endocytosis, membrane recycling, and signaling network coordinating cellular processes, structural organization and vesicular trafficking activities required for tip growth. Proteomic analysis was applied to identify Nicotiana tabacum Differentially Abundant Proteins (DAPs) after in vitro pollen tube treatment with membrane trafficking inhibitors Brefeldin A, Ikarugamycin and Wortmannin. Among roughly 360 proteins separated in two-dimensional gel electrophoresis, a total of 40 spots visibly changing between treated and control samples were identified by MALDI-TOF MS and LC–ESI–MS/MS analysis. The identified proteins were classified according to biological processes, and most proteins were related to pollen tube energy metabolism, including ammino acid synthesis and lipid metabolism, structural features of pollen tube growth as well modification and actin cytoskeleton organization, stress response, and protein degradation. In-depth analysis of proteins corresponding to energy-related pathways revealed the male gametophyte to be a reliable model of energy reservoir and dynamics.

Joel Dacks is an Associate Professor and Canada Research Chair in Evolutionary Cell Biology at the University of Alberta, a Scientific Associate at the Natural History Museum (London), and the current President of the International Society for Evolutionary Protistology. His research group studies the evolution and diversity of the eukaryotic membrane-trafficking system, from origins to potential disease therapeutics. In this interview, Joel shares some perspectives on gaining a balanced view of comparative cell biology and the importance of a constructive peer review process.

Membrane bending is a ubiquitous cellular process that is required for membrane traffic, cell motility, organelle biogenesis, and cell division. Proteins that bind to membranes using specific structural features, such as wedge-like amphipathic helices and crescentshaped scaffolds, are thought to be the primary drivers of membrane bending. However, many membrane-binding proteins have substantial regions of intrinsic disorder which lack a stable three-dimensional structure. Interestingly, many of these disordered domains have recently been found to form networks stabilized by weak, multivalent contacts, leading to assembly of protein liquid phases on membrane surfaces. Here we ask how membrane-associated protein liquids impact membrane curvature. We find that protein phase separation on the surfaces of synthetic and cell-derived membrane vesicles creates a substantial compressive stress in the plane of the membrane. This stress drives the membrane to bend inward, creating protein-lined membrane tubules. A simple mechanical model of this process accurately predicts the experimentally measured relationship between the rigidity of the membrane and the diameter of the membrane tubules. Discovery of this mechanism, which may be relevant to a broad range of cellular protrusions, illustrates that membrane remodeling is not exclusive to structured scaffolds but can also be driven by the rapidly emerging class of liquid-like protein networks that assemble at membranes.

In the early secretory pathway, the delivery of anterograde cargoes from the endoplasmic reticulum (ER) exit sites (ERES) to the Golgi apparatus is a multi-step transport process occurring via the ER-Golgi intermediate compartment (IC, also called ERGIC). While the role microtubules in ER-to-Golgi transport has been well established, how the actin cytoskeleton contributes to this process remains poorly understood. Here, we report that Arp2/3 inhibition affects the network of acetylated microtubules around the Golgi and induces the accumulation of unusually long RAB1/GM130-positive carriers around the centrosome. These long carriers are less prone to reach the Golgi apparatus, and arrival of anterograde cargoes to the Golgi is decreased upon Arp2/3 inhibition. Our data suggest that Arp2/3-dependent actin polymerization maintains a stable network of acetylated microtubules, which ensures efficient cargo trafficking at the late stage of ER to Golgi transport.